Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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83989073
TREATMENT OF IMATINIB RESISTANT LEUKEMIA USING 4-AMINOQUINOLINE-3-
CARBONITRILES HAVING MUTATION IN THE BCRABL GENE
This is a divisional application of Canadian patent application Serial No.
2,868,899 filed on
May 30, 2008.
FIELD OF THE INVENTION
[0001] The invention is directed to methods of treatment of drug-
resistant cancer. In
particular, the invention is directed to methods of treating imatinib-
resistant BcrAbl positive
leukemia. It should be understood that the expression "the invention" and the
like
encompasses the subject matter of both the parent and the present application.
BACKGROUND OF THE INVENTION
[0002] lmatinib, which is sold under the trade names Gleevac and
Glivec, has
arguably transformed the treatment of chronic myeloid leukemia by helping many
patients
achieve a nearly 90% 5-year survival rate. A subset of patients on imatinib,
which is sold
under the trade names Gleevac and Glivec, develop resistance to the drug,
often because of
bcrabl mutations in the tyrosine kinase. Treatment with imatinib has allowed
patients with
chronic myelogenous leukemia (CML) to experience a nearly 90 percent five-year
survival
rate, as the drug blocks the tyrosine kinase protein "BcrAbl," an abnormal
protein driving the
overproduction of abnormal white blood cells characteristic of leukemia.
However, many
patients have eventually developed resistance to this treatment because their
cancer cells
are able to mutate and adapt, causing their disease to relapse.
[0003] The aberrantly activated tyrosine kinase BcrAbl (the product of
bcrabl gene
and the Philadelphia Chromosome) is causally associated with Chronic
Myelogenous
Leukemia and Acute lymphocytic leukemia. Constitutive tyrosine kinase activity
of BcrAbl
promotes proliferation and survival of chronic myelogenous leukemia (CML)
cells. Inhibition
of BcrAbl tyrosine kinase activity or signaling proteins activated by BcrAbl
in CML cells blocks
proliferation and causes apoptotic cell death. The selective Abl kinase
inhibitor, STI-571
(marketed as GleevecTm), is toxic to CML cells in culture, causes regression
of CML tumors
in nude mice, and is currently used to treat CML patients. Expression of
BcrAbl in
hematopoietic stem cells promotes transformation and acts early in
leukemogenesis.
Inhibition of this kinase with STI-571 effectively controls CML in the chronic
phase of the
disease but more advanced
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1/4
patients frequently progress on STI-571 therapy. In vitro models of STI-571
resistance and clinical specimens from resistant patients demonstrated that
overexpression of other kinases or activation of distinct signaling pathways
is
associated with BcrAbl independence. Inhibition of the tyrosine kinase
activity of
BcrAbl is an effective strategy for targeting CML as demonstrated by the
clinical
efficacy of STI-571. Other molecules, including Src family kinases, play a
role in
downstream signaling from BcrAbl, and as such, are potential therapeutic
targets for
the treatment of ST1-571 -resistant disease. Src family kinases including Lyn
and Hck
have been implicated in downstream signaling from BcrAbl.
[0004] Although the selective Abl kinase inhibitor STI-571 is
efficacious and well
tolerated by most patients in chronic-stage CML, patients in accelerated and
blast
crises stages of the disease tend to be less responsive. Consequently, there
is a
need for alternative agents that are effective in late-stage disease. The
frequency of
bcr/abl mutations in CML resistant patients has increased to 90% (Hochhaus et
at.
Leukemia 2004)) from 42% Cancer Cell, Vol 2. (2), August 2002, Pages 117-
125.Imatinib
is approved as a first line therapy for the newly diagnosed CML patients.
However
resistance to imatinib due to point mutations in the bcr/abl gene is being
recognized
as a hurdle in the therapy of CML patients. Gore, Science 2001;293(5531):876-
880
and Lecoutre, Blood 2000;95(5):1758-66.
[0005] Kantarijian et at. have demonstrated that nilotinob is not
effective against
CML when patients have the amino acid mutation in BcrAbl T315I N Engl J Med.
2006 Jun 15; 354(24):2594-6.
[0006] Talpaz et at. have shown that Dasatinib in Imatinib-Resistant
Philadelphia
Chromosome-Positive Leukemias (New England J Med.2006:354:2531-2541) also
has no effect against the T315I mutation. This reference also demonstrated
that
Dasatinib can cause hematologic toxicity and edema.
[0007] Branford et al. reported that BcrAbl mutations in patients
with CML
treated with imatinib are virtually always accompanied by clinical resistance,
and
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mutations in the ATP phosphate-binding loop (P-loop) are associated with a
poor
prognosis. Blood, 1 July 2003, Vol. 102, No. 1, pp. 276-283.
[0008] U.S. Patent No. 6,297,258 discloses substituted 3-cyanoquinolines
that
are useful as antineoplastic agents and in the treatment of polycystic kidney
disease.
U.S. Patent Application No. 20050101780 discloses methods of treating
preventing
or inhibiting CML by providing to a subject a therapeutically effective amount
of SKI-
606.
[0009] U.S. Patent Publication No. 20050101780 specifically discloses the
use
of a compound having the structural formula
cH,
HN 0
C
for the treatment of CML. This compound is also known as bosutinib or SKI-606
and
has the chemical name 4-[(2,4-Dichloro-5-methoxy-phenyl)amino]-6-methoxy-743-
(4-
methyl-1-piperazinyl)propoxy]-3-quinolinecarbonitrile
[0010] Soverini et al. demonstrated the resistance to dasatinib of patients
with
F317V., J Clin Oncol. 2006 Nov 20;24(33):e51-2.
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[0011] Puttini et al. have shown that SKI-606, a novel Src-Abl inhibitor is
effective
at reducing replication of imatininb resistant CML cell lines having certain
mutations
associate with imatinib resistance. Cancer Res. 2006; 66(23):Dec 1, 2006.
SUMMARY OF THE INVENTION
[0012] It has been discovered that a significant number of imatinib resistant
patients
respond favorably to treatment with SKI-606 (4-[(2,4-Dichloro-5-methoxy-
phenyl)amino]-6-methoxy-743-(4-methy1-1-piperazinyl)propoxy]-3-
quinolinecarbonitrile).
[0013] It has also been discovered that a significant number of patients
having
known point mutations associated with resistance to imatinib respond favorably
to
treatment with SK-606. Thus, in one embodiment, the invention provides a
method of
treating a subject suffering from BcrAbl positive leukemia, wherein the
leukemia is
resistant to treatment with imatinib, the method comprising administering to
the
subject a therapeutically effective amount of a compound of the Formula:
HN
R20 CN
R¨N X¨(CH2)n
wherein:
n is 1, 2 or 3;
X is N or CH, provided that when X is N, then n is 2 or 3;
R is alkyl of from 1 to 3 carbon atoms;
R' is selected from the group consisting of 2,4-dichloro-5-methoxyphenyl; 2,4-
dichlorophenyl; 3,4,5-trimethoxyphenyl; 2-chloro-5-methoxyphenyl; 2-methyl-5-
-4-
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=
methoxyphenyl; 2,4-dimethylphenyl; 2,4-dimethy1-5-methoxyphenyl; and 2,4-
dichloro-
5-ethoxyphenyl; and
R2 is alkyl of from 1 to 2 carbon atoms;
or a pharmaceutically acceptable salt thereof.
[0014] In one
embodiment, the method comprises treating a subject suffering
from Leukemia and in some embodiments the Leukemia is selected from CML and
Acute Lymphocytic Leukemia (AML).
[0015] In some
embodiments, the invention provides a method of treatment
wherein the imatinib resistant subjects have one or more nucleic acid
mutations in
the bcrabl gene selected from the group consisting of: 10521>C; 1075T>G;
1187A>C; 1295T>C; 1457T>C; 730A>G; 742C>S; 749G>A; 757T>C; 758A>T;
763G>A; 787A>G; 817T>A; ; 944C>T; 944C>T; 949T>C; and 992A>G.
[0016] In some
embodiments, the invention provides a method of treatment
wherein the imatinib resistant subjects have one or more amino acid mutations
in
BcrAbl selected from the group consisting of: M351T;F359V; I-1396P; I432T;
F486S;
M244V; L248V; G250E; Y253H; Y253F; E255K; K263E; L273M; T3151; F317L; and
N331S.
[0017] In one
embodiment, the compositions of the present invention are
administered at a concentration selected from about 100 and about 1000 mg,
between about 200 and about 800 mg, between about 300 and about 700 mg,
between about 400 and about 600 mg and any intervals or fractions included
within
these ranges. In one
embodiment, the compounds are administered at a
concentration between 400 and 600 mg per day. In one embodiment, the compounds
are administered at a concentration at about 500 mg per day.
[0018] In another
embodiment, the invention provides a method of treating a
subject suffering from BcrAbl positive leukemia, wherein the leukemia is
resistant to
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= WO 2008/150957
PCT/US2008/065215
treatment with imatinib, the method comprising administering to the subject a
therapeutically effective amount of a compound of the Formula:
CI CI
HN OM e
R20 CN
0
R ¨ N X ¨(CH2).
wherein:
X is N or CH;
n is 3;
R2 and R are methyl;
or a pharmaceutically acceptable salt thereof.
[0019] In another embodiment, the invention provides a method of
treating a
subject suffering from BcrAbl positive leukemia, wherein the leukemia is
resistant to
treatment with imatinib, the method comprising administering to the subject a
therapeutically effective amount of a compound of the Formula:
HN e-
-
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[0020] In another embodiment, the invention provides the use of a
compound
of the Formula:
HN/ Ri
R20 CN
0
R¨N X¨(CH2),
wherein:
n is 1, 2 or 3;
X is N or CH, provided that when X is N, then n is 2 or 3;
R is alkyl of from 1 to 3 carbon atoms;
R1 is selected from the group consisting of 2,4-dichloro-5-methoxyphenyl; 2,4-
dichlorophenyl; 3,4,5-trimethoxyphenyl; 2-chloro-5-methoxyphenyl; 2-methy1-5-
methoxyphenyl; 2,4-dimethylphenyl; 2,4-dimethy1-5-methoxyphenyl; and 2,4-
dichloro-
5-ethoxyphenyl; and
R2 is alkyl of from 1 to 2 carbon atoms;
or a pharmaceutically acceptable salt thereof;
for use in the manufacture of a medicament for the treatment of imatinib
resistant
cancer.
[0021] In another embodiment, the invention provides the use of a
composition
encompassed by the formula
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83989073
HN o/cH2
=
H3C/C3
-
H3C
or a pharmaceutically acceptable salt thereof;
for use in the manufacture of a medicament for the treatment of imatinib
resistant
leukemia. The composition is also known as SKI-606 and bosutinib and has the
chemical name 4-[(2,4-Dichloro-5-methoxy-phenyl)amino]-6-methoxy-713-(4-methyl-
1-piperazinyl)propoxy]-3-guinolinecarbonitrile.
[0021A] The present disclosure includes:
- a compound of the Formula:
HN7Ri
R20 CN
0
R¨N X¨(CF12)n
wherein: n is 1, 2 or 3; X is N or CH, provided that when X is N, then n is 2
or 3; R is
alkyl of from 1 to 3 carbon atoms; R1 is selected from the group consisting of
2,4-
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= 83989073
dichloro-5-methoxyphenyl; 2,4-dichlorophenyl; 3,4,5-trimethoxyphenyl; 2-chloro-
5-
methoxyphenyl; 2-methyl-5-methoxyphenyl; 2,4-dimethylphenyl; 2,4-dimethy1-5-
methoxyphenyl; and 2,4-dichloro-5-ethoxyphenyl; and R2 is alkyl of from 1 to 2
carbon atoms; or a pharmaceutically acceptable salt thereof, in an amount
therapeutically effective for use in the treatment of BcrAbl positive
leukemia, wherein
the leukemia is resistant to imatinib, and wherein the leukemia has the
resistance-
associated amino acid mutation F317L in the bcrabl gene; and
- use of a therapeutically effective amount of 4-[(2,4-Dichloro-5-
methoxy-phenyl)amino]-6-methoxy-743-(4-methyl-1-piperazinyl)propoxy]-3-
quinolinecarbonitrile or a pharmaceutically acceptable salt thereof, for
treating a
BcrAbl positive leukemia in a subject that is resistant to imatinib, wherein
the subject
has at least one mutation in BcrAbl protein that is F317L.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] , Figure 1 shows a summary of responses hematological and
cytogenetic
responses following treatment with SKI-606.
[0023] Figure 2 shows levels of expression of bcrabl gene.
DETAILED DESCRIPTION OF THE INVENTION
General Methods
[0024] Automated complete blood counts, differential counts (with
manual
confirmation of abnormalities), bone marrow morphology, and cytogenetics are
used
to determine response to treatment.
[0025] Bone marrow morphology is used to determine the blast and
immature
myeloid cell counts in order to define disease phases.
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[0026] Standard cytogenetics are used to determine the presence of the
Philadelphia chromosome and its percent presence in marrow. Twenty or more
metaphases should be counted for this determination. FISH (Fluorescent in situ
hybridization) analysis may be used to confirm presence of BcrAbl fusion
product.
[0027] Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for BcrAbl
copy number is performed on peripheral blood.
[0028] As used herein the term "BcrAbl positive leukemia" refers to a
leukemia
that is associated with expression of the bcrabi gene.
[0029] Cytogenetic response to treatment. As used herein, a "cytogenetic
response to treatment" indicates a relative disappearance of the Philadelphia
chromosome in treated subjects as determined by a percentage of Philadelphia
chromosome positive cells present. The response can be minimal, minor, partial
or
complete. A "negative" cytogenetic response represents approximately 95.5 %
cells
positive for the Philadelphia chromosome after treatment. A "minimal response"
indicates approximately 66-95% cells positive for the Philadelphia chromosome.
A
"minor" cytogenetic response indicates 36-65% cells positive for the
Philadelphia
chromosome. A "partial" response indicates 1-35% cells positive for the PC.
complete response indicates 0% cells positive for the Philadelphia chromosome.
These figures for % positive are based on analysis of 20 metaphases (per
subject?).
A fluorescence in situ hybridization (FISH)-based assay can be used to qualify
response if insufficient metaphases are available.
[0030] Hematologic Responses to treatment. As used herein, a "hematologic
response to treatment" indicates the elimination of microscopically observed
leukemia cells in the blood.
[0031] The compounds of this invention may be used for treating,
preventing, or
inhibiting imatinib resistant leukemia. In a preferred embodiment the
compounds are
used as part of a pharmaceutical composition.
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[0032] Pharmaceutically acceptable salts are those derived from such
organic
and inorganic acids as: acetic, lactic, carboxylic, citric, cinnamic,
tartaric, succinic,
fumaric, maleic, malonic, mandelic, malic, oxalic, propionic, hydrochloric,
hydrobromic, phosphoric, nitric, sulfuric, glycolic, pyruvic, methanesulfonic,
ethanesulfonic, toluenesulfonic, salicylic, benzoic, and similarly known
acceptable
acids.
[0033] The term "alkyl" refers to the radical of saturated aliphatic
groups,
including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl
(alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl
substituted alkyl
groups. In a preferred embodiment, a straight chain or branched chain alkyl
has 3 or
fewer carbon atoms in its backbone.
[0034] Compounds may be provided orally, by intralesional,
intraperitoneal,
intramuscular or intravenous injection; infusion; liposome-mediated delivery;
topical,
nasal, anal, vaginal, sublingual, uretheral, transdermal, intrathecal, ocular
or otic
delivery. In order to obtain consistency in providing the compound of this
invention it
is preferred that a compound of the invention is in the form of a unit dose.
[0035] Suitable unit dose forms include tablets, capsules and powders in
sachets or vials. Such unit dose forms may contain from 0.1 to 1000 mg of a
compound described herein to treat imatinib resistant leukemia and preferably
from
400 to 600 mg. In another embodiment the unit dosage forms contain 500 mg of a
compound of the present invention.
[0036] In one embodiment, the daily dosage is between 400 and 600 mg per
day. In yet another embodiment, the compounds can be administered in unit
dosage
forms containing 500 mg.
[0037] The compounds of the present invention can be administered orally.
Such compounds may be administered from 1 to 6 times a day, more usually from
1
to 4 times a day. The effective amount will be known to one of skill in the
art; it will
also be dependent upon the form of the compound. One of skill in the art could
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J54-221 CA 2964162 2017-04-12
routinely perform empirical activity tests to determine the bioactivity of the
compound
in bioassays and thus determine what dosage to administer.
[0038] The compounds of the invention may be formulated with
conventional
=
excipients, such as a filler, a disintegrating agent, a binder,.a lubricant, a
flavoring
agent, a color additive, or a carrier. The carrier may be for example a
diluent, an
aerosol, a topical carrier, an aqueous solution, a nonaqueous solution or a
solid
carrier. The carrier may 'be a polymer or a toothpaste. A carrier in this
invention
encompasses any of the standard pharmaceutically accepted carriers, such as
phosphate buffered saline solution, acetate buffered saline solution, water,
emulsions
such as an oil/water emulsion or a triglyceride emulsion, various types of
wetting
= agents, tablets, coated tablets and capsules.
[0039] When provided orally or topically, .such compounds would be
provided to
a subject by delivery in different carriers. Typically, such carriers contain
exciplents
such as starch, milk, sugar, certain types of clay, gelatin, stearic acid,
talc, vegetable
fats or oils, gums, or glycols. The specific carrier would need to be selected
based
upon the desired method of delivery, for example, phosphate buffered saline
(PBS)
= could be used for intravenous or systemic delivery and vegetable fats,
creams,
salves, ointments or gels may be used for topical delivery.
= [0040] The compounds of the present invention may be
delivered together with
suitable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or
carriers
useful in treatment or prevention of neoplasm. uch compositions are liquids Or
lyophilized or otherwise dried formulations and include diluents of various
buffer
content (for example, Tris-HCl, acetate, phosphate), pH and ionic strength,
additives
such as albumins or gelatin to prevent absorption to surfaces, detergents (for
TM TM
example, TWEEN 20, TWEEN BO, PLURONIC ,F68,. bile acid salts), solubilizing
agents (for example, glycerol, polyethylene glycerol), anti-oxidants (for
example
ascorbic acid, sodium metabisulfate), preservatives (for example, thimerosal,
benzyl
alcohol, parabens), bulking substances or tonicity modifiers (for example,
lactose,
mannitol), covalent attalcliment of polymers such as polyethylene glycol,
= complexation with metal ions, or incorporation of the compound into or
onto
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=
=
= =
=
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particulate preparations of hydrogels or liposomes, micro-emulsions, micelles,
unilamellar or multilameliar vesicles, erythrocyte ghosts, or spheroblasts.
Such
compositions will influence the physical state, solubility, stability, rate of
in vivo
release, and rate of in vivo clearance of the compound or composition. The
choice of
compositions will depend on the physical and chemical properties of the
compound
capable of treating or preventing a neoplasm.
[0041] The compound of the present invention may be delivered locally via
a
capsule that allows a sustained release of the compound over a period of time.
Controlled or sustained release compositions include formulation in lipophilic
depots
(for example, fatty acids, waxes, oils).
[0042] The present invention further provides a compound of the invention
for
use as an active therapeutic substance for treating, preventing, or inhibiting
CML.
[0043] The present invention further provides a method of treating CML in
humans, which comprises administering to the infected individual an effective
amount
of a compound or a pharmaceutical composition of the invention. The dose
provided
to a patient will vary depending upon what is being administered, the purpose
of the
administration, the manner of administration, and the like. A "therapeutically
effective
amount" is an amount sufficient to cure or ameliorate symptoms of CML.
[0044] The compounds of this may be delivered alone or in combination with
other compounds used to treat CML. Such compounds include but are not limited
to
GLEEVEC, hydroxyurea, IFN-alpha, cytotoxic agents, 17-(Allylamino)-17-
demethoxygeldanamycin or derivatives thereof, or wortmannin.
[0045] The compounds of this invention were prepared from: (a)
commercially
available starting materials (b) known starting materials which can be
prepared as
described in literature procedures or (c) new intermediates described in the
schemes
and experimental procedures herein. Compounds included in this invention can
be
prepared according to the synthesis routes disclosed in U.S. Pat. Nos.
6,002,008,
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_
sO54-221 CA 2964162 2017-04-12
and 6,780,996.
[0046] Reactions are performed in a solvent appropriate to the
reagents and
materials employed and suitable for the transformation being effected. It is
understood by those skilled in the art of organic synthesis that the various
functionalities present on the molecule must be consistent with the chemical
transformations proposed. When not specified, order of synthetic steps, choice
of
protecting groups and deprotection conditions will be readily apparent to
those skilled
in the art. In addition, in some instances, substituents on the starting
materials may
be incompatible with certain reaction conditions. Restrictions pertinent to
given
= substituents will be apparent to one skilled in the art. Reactions were
run under inert
atmospheres where appropriate.
[0047] The preparation of compounds of Formula I have been reported
in the
literature, [Boschelli, D. H., et. al., J. Med. Chem., 44, 3965 (2001)],
Boschelli, D. H.,
et al., j Med. Chem., 44; 822 (2001), Boschelli, D. H., at al., Bioorg. Med.
Chem.
Lett., 13, 3797 (2003), Bdschelli, D. H., et.al., J. Med. Chem., 47, 1599
(2004), and
Ye, F. et. al., 221th National Meeting of the American Chemical Society, San
Diego,
Calif. (April, 2001)].
[0048] The present invention further provides a compound of the
invention for
use as an active therapeutia substance for treating, preventing, or inhibiting
CML in
patients that have failed to respond to treatment with imatinib.
[0049] The present invention further provides a method of treating
CML in
humans patients that have failed to respond to treatment with imatinib, which
comprises administering to the infected individual an effective amount of a
compound
or a pharmaceutical composition that is a substituted 3-cyanoquinoline. In one
embodiment, the substituted 3-cyanoquinoline is 8K606 (also known as 61<I-606
or
bosutinib. The chemical name for this compound is 4-[(2,4-Dichloro-5-methoxy-
phenypamino]-6-methoxy-743-(4-methyl-1-piperazinyl)propoxy]-3-
quinolinecarbonitrile. The dose provided to a patient will vary depending upon
what is
being administered, the purpose of the administration, the manner of
administration,
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=
=
...J54-221 CA 2964162 2017-04-12
and the like. A "therapeutically effective amount" is an amount sufficient to
cure or
ameliorate symptoms of CML In one embodiment, the method for treating a BcrAbl
positive leukemia in a subject that is resistant to imatinib comprises
administering to
the subject a therapeutically effective amount of 4-[(2,4-Dichloro-5-methoxy-
phenyl)amino]-6-methoxy-743-(4-mathyl-1-pioerazinyljpropoxy]-3-
quinolinecarbonitrile., wherein the subject has at least one mutation in
BcrAbl
selected from M351T; .F359V; H396P; I432T; F486S; M244V; 1.248V; G250E;
Y253H; Y253F; E255K; K263E; L273M; T3151; F317L; and N331S.
[0050] The
compounds of this may be delivered alone or in combination with
other compounds used to treat .CML. Such compounds include but are not limited
to
GLEEVEC, hydroxyurea, I FN-alpha cytotoxic agents, 17-(Allyiamino)-17-
demethoxygeldanamycin or derivatives thereof, or wortmannin.
[0051] The
compounds of this invention and more particularly as described
below in Examples 2 through 23 are or were prepared from: (a) commercially
available starting materials '(b) known starting materials which can be
prepared as
described in literature procedures or (c) new intermediates described in the
schemes
and experimental procedures herein. Compounds included in this invention can
be
prepared according to the synthesis routes disclosed in U.S. Pat. Nos.
6,002,008,
and 6,780,996.
EXAMPLE 1
==
[0052] Mutations
known to be associated with resistance to imatinib are located
in the bcr/abl gene are as follows, with the nucleotide position and the
nucleotide
change shown and followed in parentheses by the corresponding amino acid
change
shown in parentheses: 1052T->C (M3511); 1075T>l< (F359V); 1187A>M (H396P);
, 1295T>Y (1432T); 1457T>C (F4863); 730A>G (M244V); 742C>S (L248V);
749G>R
(G250E); 757T>C (Y253H);,758A>T (Y253F); 763G>R (E255K); 787A>R (K263E);
817T>A (L273M); 944C>T (T315I); 949T>C (F317L); and 992A>G(N331S).
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[0053] Bone marrow aspirate samples were collected from subjects who
failed
imatinib treatment for Chronic Myeloid Leukemia, prior to dosing with SKI-606.
The
baseline bcr/abl gene was sequenced and point mutations were recorded. The
patients were then dosed with SKI 606 and followed for best Cytogenetic and
confirmed Hematological responses. Doses averaged between 400 mg and 600 mg
per patient per day. It was confirmed that SKI-606 treatment resulted in
cytogenetic
or hematologic responses in patients harboring at least one of nineteen unique
point =
mutations of the bcr/abl gene. These point mutations are associated with
resistance
to treatment with imatinib: Treatment times varied from one week to greater
than a
year.
[0054] Results of treatment of imatinib resistant human subjects
having known
= BcrAbl resistance-associated mutations are shown in Table 1. A total of
66 patients
resistant to imatinib were treated with SKI-606 for times varying between one
week
and more than one year per individual subject. Of these 66 patients, 42 had
one or
more mutations known to be associated with imatinib resistance. Furthermore,
some
patients not having one of the known resistance-associated mutations also
responded favorably to treatment.
[0055] The following additional examples 2 through 23 describe compounds
useful in
the methods of the invention and are synthesized using (a) commercially
available
starting materials (b) known starting materials which can be prepared as
described in
literature procedures or (c) new intermediates described in the schemes and
experimental procedures herein. Compounds included in this invention can be
prepared according to the Synthesis routes disclosed in U.S. Pat. Nos.
6,002,008,
_
and 6,780,996. =
=
=
:
=
= I.
=
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TABLE 1
any
cytogenetic heme responder
responders responders per total best
per subjects per subjects subjects cytogenefic best heme
Mutations DNA Protein assessed
assessed assessed response response
2-Complete
heme
= response: 1-
Accelerated
thymidine (T) to methionine to 2-Ccyr, 1- to Chronic
10527>C (M351T) cytosine (C) threonine 3 out of 5 3 out of 4
4 out of 5 Pcyr phase
1- Complete
thyrnidine (T) to phenylalanine to heme
10757>G (F359V) guanine (G) valine 1 out of 1 1 out of 2 1 out
of 2 1-Ccyr 'response
1-Complete
heme
response: 1-
thymidine to Phenylalanine to Blasi CTISiS to
thymidine or Phenylalanine Chronic
1075T>K (F359[V,9) guanine and valine 1 out of 2 2 out of 3
2 out of 3 1-Micyr Phase
histidine to 1-Complete
Adenine (A) to A histidine and heme
1187A>M (H396(H,P) or C phenylalanine 1 out of 1 1 out of 1
1 out of 1 1-Ccyr response
isoleucine to 1-Complete
isoleucine and heme
12957>Y (1432[T,11) Ito T or C threonine -- 0 out of 1 -- 1 out
of 1 -- 1 out of 1 No Response response
=
1- Blast crisis
to Chronic
14571>C (F486S) T to C Phe to sense 1 out of 2 1 out of 2= 1
out of 2 1-Ctyr Phase
730A>G MUTATION
(M244V) A to G met to valine 1 out of 1 Not Evaluable 1 out of 1 -
- 1-Ccyr -- Not Evaluable
'Accelerated
phase to
met to met and Chronic
730A>R (M2441M.VD A to A or G valine -- Not Evaluable 1 out
oil . -- 1 out of 1 Not Evaluable -- phase
1-Complete
Leucine to leu heme
742C>S (L248[L,V]) C to C or G and valine 0 out of 1 1
out of 1 1 out of 1 No Response response
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TABLE 1 (CONT.)
any
cytogene tic heme responder
responders responders per total best
per subjects per subjects subjects cytogenefic best heme
Mutations DNA Protein assessed assessed
assessed response response
Glycine to 1-Complete
glutamate and heme
749G>R (G250{E,G)) G to A or G glycine Not
Evaluable 1 out of 1 1 out of 1 Not Evaluable response
tyrosine to 1-Complete
tyrosine and heme
7577>C (Y253{1-1,Y}) T to C histidine 1 out of 1 1 out of 1
1 out of 1 1-Pcyr response
1-Complete
tyrosine to heme
758A>7 (Y253F) A to 7 phenylalanine 0 out of 1 1 out
of 1 1 out of 1 No Response response
glutamate to 1-Blast crisis
glutamate and to Chronic
763G>R (E2551K,ED G to A or G lysine 1 out of 1 1 out off .. 1 out of
1 .. 1-Pcyr, .. Phase
1-Complete
lysine to lysine heme
787A>R (K263KE1) A to A or G and glutamate Not
Evaluable 1 out of 1 1 out of 1 Not Evaluable response
1-Complete
Leucine to heme
8177>A (L273M) T to A methionine 1 out of 1 1 out of 1 1 out
of 1 1-Ccyr response
1-Complete
threonine to heme
944C>T (73151) C to 7 isoleucine 1 cutoff 1 out of 6
response
threonine to
threonine and
944C>Y (7315[7.1]) C to C or 7 isoleucine 1 out of 1 0 out of
1 1 out of 1 1-Pcyr No Response
3-Complete
hems
9497>C (F317L) 7 to C Phe to lysine 1 out of 3 3 our of 4 3
out of 4 1-Micyr response
Asparagines to
992A>G(N3313) A to G serine 1 out of 1 Not Evaluable 1
out of 1 1-Ccyr Not Evaluable
Ccyr = Complete
cytogenetic response
Pcyr = Partial
crogenetic response
MiCyr= Minimal
cytogenetic response
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EXAMPLE 2
[0056] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-743-(4-methyl-1-
piperazinyl)propoxy]-3-quinolinecarbonitrile mp 116-120 C.; MS (ES) rniz
530.2,
532.2 (M+1);
EXAMPLE 3
[0057] 4-[(2,4-Dichloro-5-methoxyphenyl)amino)-743-(4-ethy1-1-
piperazinyl)propoxy]-6-methoxy-3-quinolinecarbonitrile; mp 102-104 C.; MS
(ES)
m/z 54-4.3, 546.4 (M+1);
EXAMPLE 4
[0058] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-742-(4-methyl-1-
piperazinyl)ethoxy]-3-quinolinecarbonitrile mp 165-167 C.; MS (ES) m/z 516.0,
518.2 (M+1);
EXAMPLE 5
[0059] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-742-(4-ethy1-1-
piperazinyl)ethoxy- ]-6-methoxy-3-quinolinecarbonitrile mp 101-105 C.; MS
(ES) m/z
530.4, 532.4 (M+1);
EXAMPLE 6
[0060] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[(1-
methylpiperidin-4-y1)methoxy]-3-quinolinecarbonitrile mp 200-202 C., MS 501.3
(M+H)+, Analysis for C<sub>25H</sub><sub>26C1</sub>,sub.2N<sub>40</sub><sub>3-0</sub>.8H<sub>20</sub>,
Calcd:
C, 58.21; H, 5.39; N, 10.86, Found: C, 58.19; H, 5.23; N, 10.67;
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EXAMPLE 7
[0061] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-742-(1-
methylpiperidin-- 4-yl)ethoxy}-3-quinolinecarbonitrile mp 190-191 C., MS
515.19
(M+H)+, Analysis for C<sub>26H</sub><sub>28C1</sub><sub>2N</sub><sub>40</sub><sub>3-1</sub>.0 H<sub>20</sub>,
Calcd:
C, 58.53; H, 5.67; N, 10.50, Found: C, 58.65; H, 5.57; N, 10.34
EXAMPLE 8
[0062] 4-[(2,4-Dichloro-5-methoxyphenyDamino]-6-methoxy-743-(1-
methylpiperidin¨ 4-yl)propoxy)quinoline-3-carbonitrile mp 144-145 C.; Mass
spec.
529.2 (ES+);
EXAMPLE 9
[0063] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-74(1-ethylpiperidin-4-
yOmethoxy)- -6-methoxyquinoline-3-carbonitrile mp 192-195 C.; Mass spec.
515.2
(ES+);
EXAMPLE 10
[0064] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-ethoxy-74344-
methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile mp 137-138 C., MS 542.0
(M-
H)-, Analysis for C<sub>27H</sub><sub>31C1</sub><sub>2N</sub><sub>50</sub><sub>3--0</sub>.6 H<sub>20</sub>, Calcd:
C,
58.40; H, 5.84; N, 12.61, Found: C, 58.31; H, 5.71; N, 12.43;
EXAMPLE 11
[0065] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-ethoxy-7-[(1-
methylpiperidin-
4-y-l)methoxy]quinoline-3-carbonitrile mp 182-186 C., MS 513.0 (M-H)-,
Analysis for
C<sub>26H</sub><sub>28C1</sub><sub>2N</sub><sub>40</sub><sub>3--1</sub>.4H<sub>20Calcd</sub>: C, 57.76; H, 5.74;
N,
10.36, Found: C, 57.65; H, 5.43; N, 10.15;
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EXAMPLE 12
[0066] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-ethoxy-743-(4-
ethylpiperazin-
1-yl)propoxylquinoline-3-carbonitrile mp 127-130 C., MS 558.3 (M+H)+,
Analysis for
C<sub>28H33C1</sub><sub>2N</sub><sub>50</sub><sub>3-1</sub>.5 H<sub>20</sub>, Calcd: C, 57.44; H, 6.20; N,
11.96, Found: C, 57.44; H, 6.24; N, 11.79;
EXAMPLE 13
[0067] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-ethoxy-7-[3-(1-
methylpiperidin-4-y0propoxylquinoline-3-carbonitrile mp 148-151 C. 543.2
(M+H)+,
Analysis for C<sub>28H</sub><sub>32C1</sub><sub>2N</sub><sub>40</sub><sub>-</sub> 3-1.8 H<sub>20</sub>, Calcd: C,
58.39; H, 6.23; N, 9.73, Found: C, 58.40; H, 6.16; N, 9.64;
EXAMPLE 14
[0068] 4-[(2,4-Dichloro-5-methoxyphenyl)amino]-6-ethoxy-7-[2-(4-methyl-1-
piperazinypethoxy]quinoline-3-carbonitrile mp 141-143 C., MS 530.2 (M+H)+,
Analysis for C<sub>26H</sub><sub>29C1</sub><sub>2N</sub><sub>50</sub><sub>3</sub>, Calcd: C, 58.87; H, 5.51;
N,
13.20, Found: C, 58.48; H, 5.45; N, 12.95;
EXAMPLE 15
[0069] 44(2,4-Dich)oro-5-methoxyphenyl)amino]-6-ethoxy-7-[2-(1-
methylpiperidin-4- -ypethoxy]quinoline-3-carbonitrile mp 174-175 C., MS 529.1
(M+H)+, Analysis for C<sub>27H</sub><sub>30CI</sub><sub>2N</sub><sub>40</sub><sub>3</sub>, Calcd: C, 61.25;
H,
5.71; N, 10.58, Found: C, 61.40; H, 5.84; N, 10.35;
EXAMPLE 16
[0070] 44(2,4-Dichloro-5-methoxyphenyl)amino]-6-methoxy-713-(4-propy1-1-
piperazinyl)propoxy]-3-quinolinecarbonitrile 1 C.; MS (ES) m/z 558.2, 560.2
(M+1);
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EXAMPLE 17
[0071] 4-[(2,4-dichlorophenyl)amino]-6-methoxy-7-[(1-methylpiperidin-4-
yl)methoxy- )-3-quinolinecarbonitrile mp 224-225 C., MS 469.0 (ES-);
EXAMPLE 18
[0072] 6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy)-4-[(3,4,5-
trimethoxyphenyl)amino]quinoline-3-carbonitrile mp>245 C.; HRMS (M+H)+
calculated 493.24455, found 493.24311;
EXAMPLE 19
[0073] 4-[(2-chloro-5-methoxyphenyl)amino]-6-methoxy-7-[(1-
rnethylpiperidin-4-
Arn- ethoxy)quinoline-3-carbonitrile mp 106-108 C., MS 467.2 (ES+);
EXAMPLE 20
[0074] 6-methoxy-4-[(5-methoxy-2-methylphenyl)amino]-7-[(1-methylpiperidin-
4-
y1)m- ethoxy]quinoline-3-carbonitrile mp>250 C., MS 445.2 (ES-);
EXAMPLE 21
[0075] 4-[(2,4-dimethylphenyl)amino]-6-methoxy-7-[(1-methylpiperidin-4-
yl)methoxy- ]quinoline-3-carbonitrile mp 190-191 C., MS 429.2 (ES-);
EXAMPLE 22
[0076] 6-methoxy-4-[(5-methoxy-2,4-dimethylphenyl)amino]-7-[(1-
methylpiperidin-4-yl)methoxy]quinoline-3-carbonitrile mp 160-162 C., MS 461.3
(ES+);
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EXAMPLE 23
[0077] 4-[(2,4-dichloro-5-ethoxyphenyl)amino]-6-methoxy-7-[(1-
methylpiperidin-4-
y-1)methoxy]quinoline-3-carbonitrile.
EXAMPLE 24
[0078] A group of human patients suffering from a BcrAbl positive leukemia
and
resistant to treatment with imatinib were treated with SK1-606 for time
periods ranging
between one week and more than one year.
[0079] Figure 1 shows the hematologic and cytogenetic response for patients
by
number (N) and % and differentiated between chronic and advanced leukemia.
[0080] Figure 2 shows the median bcrabl to abl gene expression ratio in
chronic
phase imatinib resistant patients treated with SKI-606.
EXAMPLE 25
[0081] Table 2 represents follow on data collected for additional
responders to
the mutations described above in Table 1 as well as responders and non-
responders
to additional bcrabl mutations.
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TABLE 2
cyto genetic cyto genetic heme
responders
responders heme responders responders per per subjects
per subjects = per subjects
subjects assessed assessed as per
Mutations assessed assessed as per TABLE 1 TABLE 1
M351T 4 out of 6 5 out of 5 3 out of 5 3 out of 4
F359V 3 out of 4 5 out of 5 2 out of 3 3 out of 5
H396P I out of 1 1 out of 1 1 out of 1 1 out of 1
1432T 0 out of 1 I out of 1 0 out of 1 1 out of 1
,--,--; F486S 1 out of 2 1 out of 2 1 out of 2 1 out of 2
4244V 4 out of 4 4 out of 4 1 out of 1 1 out of 1
L248V 1 out of 3 2 out of 2 0 out of 1 1 out of 1
G250E 0 out of 1 2 out of 2 Not Evaluable 1
out of 1
Y253{H,F) 3 out of 3 3 out of 3 1 out of 2 2 out of 2
E255K 2 out of 2 , 2 out of 3 1 out of 1 1 out of
1
K263E 1 out of 1 1 out of / Not Evaluable 1
out of 1
T3151 3 out of 3 5 out of 9 1 out of 1 1 out of 2
F317L 1 out of 6 7 out of 8 , 1 out of 3 3
our of 4
N331 S 1 out of I Not Evaluable 1 out of 1
Not Evaluable
, 1384P 0 out of I Gout oil
V299L Gout of 1 Gout oil
E453K lout of 1 lout of 1
F359/ 1 out of 1 1 out of 1
E355G Gout of 1 lout of 1
G321R Gout of 1 1 out of 1
H396R 0 out of 2 1 out of 1
F31 1L Not Evaluable Not Evaluable
E255V Not Evaluable 1 out of 2
L273M 1 out of 1 1 out of I 1 out of 1 1 out of 1
T277A lout of 1 lout oil
E286G lout of 1 lout oil
1387V Gout of 1 , lout of 1
Q252H Not Evaluable Not Evaluable
Y230H Gout of 1 lout of 1
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